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Heterologous expression of bacteriocin E-760 in Chlamydomonas reinhardtii and functional analysis

Phyton-International Journal of Experimental Botany 2019年第1期88卷 25-35页

作者： Quezada-Rivera JJ RE Soria-Guerra FS Pérez-Juárez L Martínez-González SE Valdés-Rodríguez NL Vasco-Méndez jf morales-domínguez Departamento de Química Universidad Autónoma de AguascalientesUniversidad No.940Ciudad UniversitariaC.P.20131AguascalientesAgs.México Facultad de Ciencias Químicas Universidad Autónoma de San Luis PotosíZona Universitaria PonienteC.P.78290San Luis PotosíS.L.P.México Departamento de Biotecnología y Bioquímica Centro de Investigación y de Estudios Avanzados del Instituto Politécnico NacionalCinvestav-Unidad Irapuato.Km.9.6 Libramiento Norte Carr.Irapuato-León 36824 IrapuatoGto.México

The use of antimicrobial peptides(AMPs)synthesized by bacteria(bacteriocins)is an alternative for combating multidrug resistant bacterial strains and their production by recombinant route is a viable option for their mass production.The bacteriocin E-760 isolated from the genus Enterococcus sp.has been shown to possess inhibitory activity against Gram-negative and Gram-positive bacteria.In this study,the expression of a chimeric protein coding for E-760 in the nucleus of C.reinhardtii was evaluated,as well as,its antibacterial activity.The synthetic gene E-760S was inserted into the genome of C.reinhardtii using Agrobacterium tumefaciens.A transgenic line was identified in TAP medium with hygromycin and also by PCR.The increment in the culture medium temperature of the transgenic strain at 35°C for 10 minutes,increased the production level of the recombinant protein from 0.14(Noninduced culture,NIC)to 0.36%(Induced culture,IC)of total soluble proteins(TSP);this was quantified by an ELISA assay.Recombinant E-760 possesses activity against Staphylococcus aureus in 0.34 U log,Streptococcus agalactiae in 0.48 U log,Enterococcus faecium in 0.36 U log,Pseudomonas aeruginosa in 2 U log and for Klebsiella pneumoniae,the activity was 0.07 U log.These results demonstrate that the nucleus transformation of C.reinhardtii can function as a stable expression platform for the production of the synthetic gene E-760 and it can potentially be used as an antibacterial agent.

关键词： Heterologous expression Antibacterial activity Bacteriocin E-760 Chlamydomonas reinhardtii Log inactivation Nuclear transformation

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Benzyl Amino Purine and Gibberellic Acid Coupled to Nitrogen-Limited Stress Induce Fatty Acids,Biomass Accumulation,and Gene Expression in Scenedesmus Obliquus

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Phyton-International Journal of Experimental Botany 2021年第2期90卷 515-531页

作者： Hans Christian Correa-Aguado Gloria Viviana Cerrillo-Rojas Alejandro Rocha-Uribe Ruth Elena Soria-Guerra JoséFrancisco morales-domínguez Departamento de Química Centro de Ciencias BásicasUniversidad Autónoma de AguascalientesAguascalientes20131México Facultad de Ciencias Químicas Universidad Autónoma de San Luis PotosíSan Luis Potosí78210México Instituto Politécnico Nacional Unidad Profesional Interdisciplinaria de Ingeniería Campus ZacatecasBlvd.del Bote 202Cerro del Gato Ejido La EscondidaCol.Ciudad AdministrativaZacatecasZAC 98160Mexico

The need for renewable energy sources makes microalgae an essential feedstock for biofuels production.The molecular aspects and the response to nitrogen(N)-limited conditions with a phytohormone stimulus in microalgae have been slightly explored.In this work,Scenedesmus obliquus was used as a study model to analyze the effect of benzyl amino purine(BAP)and gibberellic acid(GA)coupled to nitrogen limitation on cell growth,biomass and fatty acids.The selected 10^(-5)M BAP increased the biomass by 1.44-fold,and 10^(-6)M GA by 1.35-fold.The total lipids also increased by 2.8 and 1.11-fold,respectively.The 10^(-5)M BAP and 10^(-6)M GA addition to S.obliquus cultures at different initial nitrogen percentages(N-0,N-25,and N-50)showed a significant increase in cell growth and biomass productivity compared to the unstimulated cultures.BAP N-0 and GA N-0 produced the highest lipid yields with 55%and 50%,respectively.The lipid profile analysis revealed an increase,particularly in C18:1 and C16:0 fatty acids.Gene expression analysis showed an over-expression of acyl carrying protein(ACP),stearoyl-ACP desaturase(SAD),fatty acid acyl-ACP thioesterase(FATA),and diacylglycerol acyltransferase(DGAT)genes,which were mainly induced by nitrogen limitation.Furthermore,BAP and GA produced a significant over-expression on these genes in the N-replete cultures.This study shows that BAP and GA,coupled to N limitation stress,can be used to increase the biomass and lipid production in S.obliquus for sustainable biofuels.

关键词： Biodiesel gene expression lipids microalgae nitrogen deficiency phytohormones

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In Vitro Propagation of Agave guiengola Gentry Using Semisolid Medium and Temporary Immersion Bioreactors

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Phyton-International Journal of Experimental Botany 2021年第3期90卷 1003-1013页

作者： Lucía Isabel Chávez-Ortiz JoséFrancisco morales-domínguez Araceli Rodríguez-Sahagún Eugenio Pérez-Molphe-Balch Departamento de Química Centro de Ciencias BásicasUniversidad Autónoma de AguascalientesAguascalientes20131México Laboratorio de Biología Molecular Vegetal Centro Universitario de la CiénegaUniversidad de GuadalajaraOcotlán47820México

Agave guiengola Gentry is an endemic plant from a very small locality in Oaxaca,Mexico.Its conservation status is fragile and can rapidly worsen.Because of its scarcity,this agave has been used solely for ornamental purposes,but it could have other uses if more plants were available.In vitro propagation by enhanced axillary sprouting from stem segments was attained using Murashige and Skoog Basal Medium(MS)as well as basal medium supplemented with cytokinins 6-Benzylaminopurine(BA)or 6-(γ,γ-Dimethylallylamino)purine(2iP).The best treatment for shoot induction in semisolid medium consisted in MS supplemented with 2 mg l^(–1) BA,obtaining a mean of 3.7 shoots per explant.Other interesting responses were observed,such as nodular callus induction using combinations of BA and 2,4-Dichlorophenoxyacetic acid(2,4-D);root induction without Plant Growth Regulators(PGR);and generation of shoot clusters.These clusters constituted an excellent explant for micropropagation in temporary immersion bioreactors,obtaining a propagation rate of 43 shoots per explant with 1 min immersion and 6 h immersion frequencies.All new plants rooted and survived the transfer to soil.This study developed an in vitro propagation scheme to produce individuals that can be used either for reforestation,economical purposes,or to carry out studies in this species to assess its full potential,avoiding exploitation from wild plants.

关键词： Agavaceae temporary immersion systems callus induction micropropagation

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Effects of organic chromium supplementation to finishing lambs diet on growth performance, carcass characteristics and meat quality

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Journal of Integrative Agriculture 2015年第3期14卷 567-574页

作者： Laura Moreno-Camarena Ignacio domínguez-Vara José Bórquez-Gastelum Juan Sánchez-Torres Juan Pinos-Rodríguez Antonia Mariezcurrena-Berasain Ernesto morales-Almaráz Abdelfattah Z M Salem Faculty of Veterinary Medicine and Animal Science Autonomous University of the State of Mexico Center of Biosciences University Autonomous of San Luis PotosiPalma of the Cruz Soledad of Graciano Sánchez 78321Mexico

The objective of this study was to evaluate supplemental organic chromium（Cr） to finishing lambs on growth performance, carcass characteristics, and meat quality. Eighteen Suffolk lambs（age（4.5±0.2） mon;（25.8±3.6） kg body weight（BW）） were randomly assigned to three levels of supplemental organic Cr（0.0, 0.2 and 0.4 mg kg^–1 dry matter（DM）） in a complete random design. Growth performance was evaluated for 70 d, and then lambs were slaughtered to study carcass characteristics and chemical composition of meat. Orthogonal contrasts were performed（contrast one-average level 0.2 ppm Cr vs. average level 0.4 ppm Cr; contrast two-level 0 vs. average levels（0.2＋0.4） ppm Cr）. Orthogonal polynomials were used to estimate the linear and quadratic effects of Cr concentrations. Growth and carcass performance were not affected by supplemental organic Cr. Muscle conformation and leg perimeter linearly increased（P〈0.05） as organic Cr level increased in the diet. Kidney fat decreased linearly（P〈0.05） as supplemental Cr increased. In Longissimus dorsi（LD）, the ash content decreased linearly, and shear force（kg cm^–2） increased（P〈0.05） as organic Cr level increased in the diet. It is concluded that organic Cr did not affect growth performance, but it improved positively the muscle conformation, reduced kidney fat, whereas in LD there was an increment in shear force in finishing carcass lambs.

关键词： carcass,finishing lambs,Longissimus dorsi,organic chromium

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Probable benign paroxysmal positional vertigo, spontaneously resolved:Incidence in medical practice, patients’ characteristics and the natural course

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Journal of Otology 2019年第3期14卷 111-116页

作者： M.G.Alvarez-Morujo de Sande R. Gonzalez-Aguado G. Guerra-Jimenez E. domenech-Vadillo H. Galera-Ruiz E. Figuerola-Massana A. Ramos-Macías C. morales-Angulo A.J. Martín-Mateos E. domínguez-Duran Hospital Universitario Puerta del Mar Cadiz Spain Hospital Universitario Marques de Valdecilla Santander Spain Complejo Hospitalario Universitario Insular Materno-Infantil Las Palmas de Gran Canaria Spain Hospital Universitari Joan XXIII Tarragona Spain Hospital Infanta Luisa Sevilla Spain

Background: Probable benign paroxysmal positional vertigo, spontaneously resolved (pBPPVsr), is a variant of benign paroxysmal positional vertigo (BPPV) in which there is no observable nystagmus and no vertigo with any positional maneuver. Objectives: To calculate the incidence pBPPVsr, compare the characteristics of the patients with pBPPVsr and BPPV not spontaneously resolved and describe the spontaneous resolution in the natural course of BPPV. Methods: Multicenter prospective descriptive study. During a one-year period, all patients with suspected BPPV that presented to the Neurotology Units of five participating centers were recruited. The incidence of pBPPVsr was calculated as a percentage of the total number of patients with BPPV. The prevalence of several variables was compared between pBPPVsr and BPPV not spontaneously resolved. The timing of spontaneous resolution was estimated using Kaplan-Meier curves. Results: 457 patients met the inclusion criteria. The incidence of pBPPVsr was 33.5%. It was significantly higher in males, in patients with normal bone mass and in patients who were not taking sulpiride. A rate of 18% of spontaneous resolution after the first month and 51% after the first year was found. This percentage did not change in a significant way after this moment. The curves for males, patients under 50 and patients with normal blood pressure decreased significantly faster. Conclusions: In our serie, BPPV spontaneously resolved in half of the patients with BPPV during the first year. This seemed to occur more commonly in males and could have been hindered by sulpiride intake, osteoporosis, advanced age and high blood pressure.

关键词： Benign paroxysmal positional vertigo Probable benign paroxysmal positional vertigo, spontaneously resolved Osteoporosis Sulpiride Hypertension

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WJD 5^(th) Anniversary Special Issues(2): Type 2 diabetes Inflammation in diabetic kidney disease

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World Journal of Diabetes 2014年第4期5卷 431-443页

作者： Patricia M García-García María A Getino-Melian Virginia domínguez-Pimentel Juan F Navarro-Gonzalez Nephrology Service and Research Unit University Hospital Nuestra Se?ora de Candelaria 38010 Santa Cruz de Tenerife Spain Spanish National Coordinator of GEENDIAB (Grupo Espa ol para el Estudio de la Nefropatía Di-abética) (RedInRen 12/0021/0019 Instituto de Salud Carlos IIIMinisterio de Economía y Competitividad) 38010 Santa Cruz de Tenerife Spain

Diabetes mellitus entails significant health problems worldwide.The pathogenesis of diabetes is multifactorial,resulting from interactions of both genetic and environmental factors that trigger a complex network of pathophysiological events,with metabolic and hemodynamic alterations.In this context,inflammation has emerged as a key pathophysiology mechanism.New pathogenic pathways will provide targets for prevention or future treatments.This review will focus on the implications of inflammation in diabetes mellitus,with special attention to inflammatory cytokines.

关键词： Diabetes Diabetic nephropathy Diabetic kidney disease Inflammation Cytokines Oxidative stress

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Innovate combination of sevoflurane dilution in dimethyl sulfoxide: A stability study by gas chromatography and nuclear magnetic resonance

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World Journal of Pharmacology 2016年第3期5卷 59-67页

作者： F Dámaso Fernández-Ginés Sergio García-Mu?oz Héctor Mateo-Carrasco Miguel ángel Rincón-Cervera Manuel Corti?as-Sáenz José Antonio morales-Molina Carmen Fernández-Sánchez Juan Miguel Expósito-López Ignacio Rodríguez-García Department of Pharmacy Torrecárdenas Hospital University of Almería Aseptic Services Unit Department of Pharmacy Northampton General Hospital Institute of Nutrition and Food Technology University of Chile Department of Anesthesiology and Pain Management Torrecárdenas Hospital Department of Pharmacy Agencia Pública Empresarial Sanitaria Hospital de Poniente Department of Chemistry and Physics University of Almería

AIM To investigate physicochemical stability of sevoflurane in dimethyl sulfoxide using gas chromatography with a flame ionization detector and nuclear magnetic resonance(NMR).METHODS Undiluted sevoflurane, plus dilutions 1:2, 1:5, 1:10, 1:25, and 1:50 in dimethyl sulfoxide were prepared in a vertical laminar flow cabinet class Ⅱ type B and stored at different temperatures(23 ℃, 6 ℃, and-10 ℃) for 45 d. Sterile 1 mL polypropylene amber syringes to minimize light degradation, caps and needles were used. The presence of sevoflurane and its degradation products in the samples was determined by gas chromatography with flame ionization detector(260 ℃, 40 min), and by ~1H, ^(19) F, and proton-decoupled ^(19) F nuclear magnetic resonance.RESULTS The gas chromatography analysis showed sevoflurane and dimethyl sulfoxide(DMSO) retention times were 2.7 and 13.0 min, respectively. Pure DMSO injection into the column resulted in two additional peaks at 2.1 and 2.8 min. The same sevoflurane peak at 2.7 min was observed in all the dilutions at-10 ℃, 4 ℃ and 25 ℃. The NMR spectra showed signals consistent with the sevoflurane structure in all the dilutions at-10 ℃, 4 ℃ and 25 ℃. In the ~1H spectrum, two signals corresponding to the sevoflurane molecule were observed at 5.12 and 4.16 parts per million(ppm5). In the ^(19)F-NMR spectrum, two signals were observed at-76.77 ppm and-157.13 ppm. In the ^(19)F NMR CPD, two signals were observed at-76.77 ppm and-157.13 ppm. The first one showed a doublet(jf-F = 3.1 Hz) which integrated by six fluorine nuclei from the hexafluoro-isopropyl group. The second signal was integrated by a fluorine atom and showed a septuplet(jf-F = 3.1 Hz).CONCLUSION This study shows that different concentrations of sevoflurane in dimethyl sulfoxide retain their chemical composition after exposure to different temperatures for a period of 45 d.

关键词：七氟醚分子药物治疗治疗方法临床分析

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